Team:Goettingen/week8-2

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Revision as of 13:20, 18 September 2012

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Chemical transformartion of the BioBrick plasmid pSB1C3 into E. coli (DH10B)

Experiment:
In order to gain further plasmid material of the new vector pSB1C3 was transformed into E. coli (DH10B) as described in the protocol.
Observations & Results:
The transformation was not successful. No colonies could be observed at the plates. Since this vector has a chloramphenicol resistance instead of ampicillin as the usually used pUC18 we suggest that the antibiotics concentration in the agar was simply to high.

V06_19_1 Repetition of the chemical transformartion of the BioBrick plasmid pSB1C3 into E. coli (DH10B)

Experiment:
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
Observations & Results:
The transformation was successful.Due to the possesion of a gene encoding for RFP the developed colonied featured a red color. On the negative control no colonied were observed.

V06_19_2 Repetition of the chemical transformartion of the araC-promoter into E. coli (DH10B)

Experiment:
Since the last time the fact that the araC-promoter I0500 (14N) is inducible by ITPG was not considered, this time the substance was added to the LB-Medium (end concentration 100 µM). This is the only deogation from the standard transformation protocol.
Observations & Results:
The transformation was again not successful. Only the positive control showed any growth.

Preparation of over night cultures

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 <ul>
 <li>Experiment: <br>
-In order to gain further plasmid material of the new vector pSB1C3 was transformed into <i>E. coli</i>(DH10B) as described in the protocol. <br>
+In order to gain further plasmid material of the new vector pSB1C3 was transformed into <i>E. coli</i> (DH10B) as described in the protocol. <br>
 <li>Observations & Results: <br>
 The transformation was not successful. No colonies could be observed at the plates. Since this vector has a chloramphenicol resistance instead of ampicillin as the usually used pUC18 we suggest that the antibiotics concentration in the agar was simply to high.</li>